title: Activation of an immunoregulatory and antiviral gene expression program in poly(I:C)-transfected human neutrophils.
abstract: Neutrophils, historically known for their involvement in acute inflammation, are also targets for infection by many different DNA and RNA viruses. However, the mechanisms by which they recognize and respond to viral components are poorly understood. Polyinosinic:polycytidylic acid (poly(I:C)) is a synthetic mimetic of viral dsRNA that is known to interact either with endosomal TLR3 (not expressed by human neutrophils) or with cytoplasmic RNA helicases such as melanoma differentiation-associated gene 5 (MDA5) and retinoic acid-inducible gene I (RIG-I). In this study, we report that intracellularly administered poly(I:C) stimulates human neutrophils to specifically express elevated mRNA levels encoding type I IFNs, immunoregulatory cytokines, and chemokines, such as TNF-alpha, IL-12p40, CXCL10, CXCL8, CCL4, and CCL20, as well as classical IFN-responsive genes (IRG), including IFIT1 (IFN-induced protein with tetratricopeptide repeats 1)/IFN-stimulated gene (ISG)56, G1P2/ISG15, PKR (dsRNA-dependent protein kinase), and IFN-regulatory factor (IRF)7. Investigations into the mechanisms whereby transfected poly(I:C) promotes gene expression in neutrophils uncovered a crucial involvement of the MAPK-, PKR-, NF-kappaB-, and TANK (TNF receptor-associated NF-kappaB kinase)-binding kinase (TBK1)/IRF3-signaling transduction pathways, as illustrated by the use of specific pharmacological inhibitors. Consistent with the requirement of the cytoplasmic dsRNA pathway for antiviral signaling, human neutrophils were found to constitutively express significant levels of both MDA5 and RIG-I, but not TLR3. Accordingly, neutrophils isolated from MDA5-deficient mice had a partial impairment in the production of IFN-beta and TNF-alpha upon infection with encephalomyocarditis virus. Taken together, our data demonstrate that neutrophils are able to activate antiviral responses via helicase recognition, thus acting at the frontline of immunity against viruses.
authors.affiliation: Department of Pathology, University of Verona, Verona, Italy.
authors.forename: Nicola
authors.initials: N
authors.lastname: Tamassia
authors.forename: Vincent
authors.initials: V
authors.lastname: Le Moigne
authors.forename: Marzia
authors.initials: M
authors.lastname: Rossato
authors.forename: Marta
authors.initials: M
authors.lastname: Donini
authors.forename: Stephen
authors.initials: S
authors.lastname: McCartney
authors.forename: Federica
authors.initials: F
authors.lastname: Calzetti
authors.forename: Marco
authors.initials: M
authors.lastname: Colonna
authors.forename: Flavia
authors.initials: F
authors.lastname: Bazzoni
authors.forename: Marco A
authors.initials: MA
authors.lastname: Cassatella
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D015967:Gene Expression Regulation, Viral / Q000276:immunology*; D005814:Genes, Viral / Q000276:immunology*; D006801:Humans; D050835:Interferon Regulatory Factors / Q000235:genetics; D016899:Interferon-beta / Q000235:genetics; D007424:Intracellular Fluid / Q000201:enzymology / Q000276:immunology / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D018375:Neutrophil Activation / Q000235:genetics / Q000276:immunology*; D009504:Neutrophils / Q000201:enzymology / Q000276:immunology* / Q000821:virology*; D011070:Poly I-C / Q000008:administration & dosage / Q000235:genetics*; D020365:RNA Helicases / Q000502:physiology; D015398:Signal Transduction / Q000235:genetics* / Q000276:immunology*; D014162:Transfection
chemical_list: D050835:Interferon Regulatory Factors; D016899:Interferon-beta; D020365:RNA Helicases; D011070:Poly I-C